用于在活体大脑中特异性检测苯丙胺慢性暴露的基于DNA的磁共振成像探针。
DNA-based MRI probes for specific detection of chronic exposure to amphetamine in living brains.
作者信息
Liu Christina H, Ren Jia Q, Yang Jinsheng, Liu Charng-ming, Mandeville Joseph B, Rosen Bruce R, Bhide Pradeep G, Yanagawa Yuchio, Liu Philip K
机构信息
Laboratory for Gene Transcript Targeting, Imaging and Repair, Massachusetts General Hospital, Charlestown, Massachusetts 02129, USA.
出版信息
J Neurosci. 2009 Aug 26;29(34):10663-70. doi: 10.1523/JNEUROSCI.2167-09.2009.
We designed phosphorothioate-modified DNA probes linked to superparamagnetic iron oxide nanoparticles (SPION) for in vivo magnetic resonance imaging (MRI) of fosB and Delta fosB mRNA after amphetamine (AMPH) exposure in mice. Specificity of both the fosB and Delta fosB probes was verified by in vitro reverse transcriptase-PCR amplification to a single fragment of total cDNA obtained from acutely AMPH-exposed mouse brains. We confirmed time-dependent uptake and retention profiles of both probes in neurons of GAD67-green fluorescent protein knock-in mice. MRI signal of SPION-labeled fosB probe delivered via intracerebroventricular route was elevated in both acutely and chronically AMPH-exposed mice; the signal was suppressed by dopaminergic receptor antagonist pretreatment. SPION-labeled Delta fosB probe signal elevation occurred only in chronically AMPH-exposed mice. The in vivo target specificity of these probes permits reliable MRI visualization of AMPH-induced differential elevations of fosB and Delta fosB mRNA in living brains.
我们设计了与超顺磁性氧化铁纳米颗粒（SPION）相连的硫代磷酸酯修饰的DNA探针，用于在小鼠安非他明（AMPH）暴露后对fosB和ΔfosB mRNA进行体内磁共振成像（MRI）。通过体外逆转录聚合酶链反应（RT-PCR）扩增，从急性暴露于AMPH的小鼠大脑中获得的总cDNA的单个片段，验证了fosB和ΔfosB探针的特异性。我们证实了两种探针在GAD67-绿色荧光蛋白基因敲入小鼠神经元中的时间依赖性摄取和保留情况。通过脑室内途径递送的SPION标记的fosB探针的MRI信号在急性和慢性暴露于AMPH的小鼠中均升高；该信号被多巴胺能受体拮抗剂预处理所抑制。SPION标记的ΔfosB探针信号升高仅发生在慢性暴露于AMPH的小鼠中。这些探针的体内靶标特异性允许在活体大脑中可靠地进行MRI可视化AMPH诱导的fosB和ΔfosB mRNA的差异升高。
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